Study of antioxidant activity and free radical scavenging power of Physalis alkekengi flower extract

According to the undesirable effects of many chemical preservatives in food products such as oilseed extraction industry, the possibility of substituting these materials with effective compounds of herbal plants have been considered by the researchers. In this study, at first, the Physalis Alkekengi flower extract was extracted by using maceration method with methanol. The Phenolic compounds and the amount of free radical scavenging activity of the flower extract were investigated in different concentrations (200, 400, 600, 800 and 1000 ppm), respectively by Folin–Ciocalteu method and DPPH test and were compared with the synthetic antioxidant activity (BHT) at 200 ppm. The results showed a significant difference between different concentrations of Physalis Alkekengi flower extract, in terms of the rate of Phenolic compounds and free radical scavenging activity of flower extract (p < 0.05). In general, the results of this study showed that the methanol extract of Physalis Alkekengi flower, as a source of cheap and available natural antioxidant, after conducting supplementary experiments can be used in food industry.Keywords: Physalis Alkekengi flower, Free radical scavenging power, Natural antioxidants,Phenolic compounds

Stable Knockdown of Adenosine Kinase by Lentiviral Anti-ADK miR-shRNAs in Wharton�s Jelly Stem Cells

Objective: In this study, we describe an efficient approach for stable knockdown of adenosine kinase (ADK) using lentiviral system, in an astrocytoma cell line and in human Wharton�s jelly mesenchymal stem cells (hWJMSCs). These sources of stem cells besides having multilineage differentiation potential and immunomodulatory activities, are easily available in unlimited numbers, do not raise ethical concerns and are attractive for gene manipulation and cell-based gene therapy. Materials and Methods: In this experimental study, we targeted adenosine kinase mRNA at 3' and performed coding sequences using eight miR-based expressing cassettes of anti-ADK short hairpin RNA (shRNAs). First, these cassettes with scrambled control sequences were cloned into expressing lentiviral pGIPZ vector. Quantitative real time-polymerase chain reaction (qRT-PCR) was used to screen multi-cassettes anti-ADK miR-shRNAs in stably transduced U-251 MG cell line and measuring ADK gene expression at mRNA level. Extracted WJMSCs were characterized using flow cytometry for expressing mesenchymal specific marker (CD44+) and lack of expression of hematopoietic lineage marker (CD45-). Then, the lentiviral vector that expressed the most efficient anti-ADK miR-shRNA, was employed to stably transduce WJMSCs. Results: Transfection of anti-ADK miR-shRNAs in HEK293T cells using CaPO4 method showed high efficiency. We successfully transduced U-251 cell line by recombinant lentiviruses and screened eight cassettes of anti-ADK miR-shRNAs in stably transduced U-251 MG cell line by qRT-PCR. RNAi-mediated down-regulation of ADK by lentiviral system indicated up to 95 down-regulation of ADK. Following lentiviral transduction of WJMSCs with anti-ADK miR-shRNA expression cassette, we also implicated, down-regulation of ADK up to 95 by qRT-PCR and confirmed it by western blot analysis at the protein level. Conclusion: Our findings indicate efficient usage of shRNA cassette for ADK knockdown. Engineered WJMSCs with genome editing methods like CRISPR/cas9 or more safe viral systems such as adeno-associated vectors (AAV) might be an attractive source in cell-based gene therapy and may have therapeutic potential for epilepsy

Stable Knockdown of Adenosine Kinase by Lentiviral Anti-ADK miR-shRNAs in Wharton�s Jelly Stem Cells

Objective: In this study, we describe an efficient approach for stable knockdown of adenosine kinase (ADK) using lentiviral system, in an astrocytoma cell line and in human Wharton�s jelly mesenchymal stem cells (hWJMSCs). These sources of stem cells besides having multilineage differentiation potential and immunomodulatory activities, are easily available in unlimited numbers, do not raise ethical concerns and are attractive for gene manipulation and cell-based gene therapy. Materials and Methods: In this experimental study, we targeted adenosine kinase mRNA at 3' and performed coding sequences using eight miR-based expressing cassettes of anti-ADK short hairpin RNA (shRNAs). First, these cassettes with scrambled control sequences were cloned into expressing lentiviral pGIPZ vector. Quantitative real time-polymerase chain reaction (qRT-PCR) was used to screen multi-cassettes anti-ADK miR-shRNAs in stably transduced U-251 MG cell line and measuring ADK gene expression at mRNA level. Extracted WJMSCs were characterized using flow cytometry for expressing mesenchymal specific marker (CD44+) and lack of expression of hematopoietic lineage marker (CD45-). Then, the lentiviral vector that expressed the most efficient anti-ADK miR-shRNA, was employed to stably transduce WJMSCs. Results: Transfection of anti-ADK miR-shRNAs in HEK293T cells using CaPO4 method showed high efficiency. We successfully transduced U-251 cell line by recombinant lentiviruses and screened eight cassettes of anti-ADK miR-shRNAs in stably transduced U-251 MG cell line by qRT-PCR. RNAi-mediated down-regulation of ADK by lentiviral system indicated up to 95 down-regulation of ADK. Following lentiviral transduction of WJMSCs with anti-ADK miR-shRNA expression cassette, we also implicated, down-regulation of ADK up to 95 by qRT-PCR and confirmed it by western blot analysis at the protein level. Conclusion: Our findings indicate efficient usage of shRNA cassette for ADK knockdown. Engineered WJMSCs with genome editing methods like CRISPR/cas9 or more safe viral systems such as adeno-associated vectors (AAV) might be an attractive source in cell-based gene therapy and may have therapeutic potential for epilepsy

Selective β2 adrenergic agonist increases Cx43 and miR-451 expression via cAMP-Epac

It has been demonstrated that connexin 43 (Cx43) and microRNAs have significant roles in glioma. Cyclic adenosine monophosphate (cAMP) is suggested to be a regulator of connexins and microRNAs. However, it remains elusive whether cAMP and exchange protein directly activated by cAMP (Epac2), have a regulatory effect on Cx43 and microRNA-451 (miR-451) in astrocytoma cells. We treated 1321N1 astrocytoma cells with a selective β2 adrenergic agonist and a selective Epac activator with and without adenyl cyclase and protein kinase A inhibition. Cx43 and miR-451 expression were measured. Next, we evaluated the effect of miR-451 overexpression on Cx43 expression. Cell proliferation was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results demonstrated that cAMPE-pac2 increased Cx43 and miR-451 expression. However, the alteration of miR-451 expression required a higher dose of drugs. Overexpression of miR-451 had no significant effect on Cx43 expression. The MTT assay showed that cAMP-Epac stimulation and miR-451 overexpression had a synergic inhibitory effect on cell proliferation. These findings may expand our understanding of the molecular biology of glioma and provide new potential therapeutic targets

A Cohort Location Model of household sorting in US metropolitan regions

In this paper we propose a household sorting model for the 50 largest US metropolitan regions and evaluate the model using 2010 Census data. To approximate residential locations for household cohorts, we specify a Cohort Location Model (CLM) built upon two principle assumptions about housing consumption and metropolitan development/land use patterns. According to our model, the expected distance from the household’s residential location to the city centre(s) increases with the age of the householder (as a proxy for changes in housing career over life span). The CLM provides a flexible housing-based explanation for household sorting patterns in US metropolitan regions. Results from our analysis on US metropolitan regions show that households headed by individuals under the age of 35 are the most common cohort in centrally located areas. We also found that households over 35 are most prevalent in peripheral locations, but their sorting was not statistically different across space

Data for: kluster: An Efficient Scalable Procedure for Approximating the Number of Clusters in Unsupervised Learning

182 simulated datasets (first set contains small datasets and second set contains large datasets) with different cluster compositions – i.e., different number clusters and separation values – generated using clusterGeneration package in R. Each set of simulation datasets consists of 91 datasets in comma separated values (csv) format (total of 182 csv files) with 3-15 clusters and 0.1 to 0.7 separation values. Separation values can range between (−0.999, 0.999), where a higher separation value indicates cluster structure with more separable clusters. Size of the dataset, number of clusters, and separation value of the clusters in the dataset is printed in file name. size_X_n_Y_sepval_Z.csv: Size of the dataset = X number of clusters in the dataset = Y separation value of the clusters in the dataset =

Age matters: ageing and household energy demand in the United States

Age is an important proxy for many life course trajectories, and has complex and understudied relationships with energy consumption. We evaluated the presence and the shape of an age-energy consumption profile in the U.S. residential sector, using household-level data from four waves of the Residential Energy Consumption Survey (RECS) in 1987, 1990, 2005, and 2009. We constructed pseudo-cohorts from Bayesian generalized linear model estimates to create micro-profiles for energy consumption across the life course. Overall, we found that residential energy consumption increases over the life course. Much of the increase in energy consumption is due to housing size. Variations in the age-energy consumption micro-profiles can be described by concave and convex functions that transform from one to another across the life course. We conclude with a demographic perspective on the future of residential energy demand in the U.S. The confluence of demographic and climatic changes will likely cause an amplification of effects, challenging the supply and demand of energy for the older population

simulated data for clustering experiments

182 simulated datasets (first set contains small datasets and second set contains large datasets) with different cluster compositions – i.e., different number clusters and separation values – generated using clusterGeneration package in R. Each set of simulation datasets consists of 91 datasets in comma separated values (csv) format (total of 182 csv files) with 3-15 clusters and 0.1 to 0.7 separation values. Separation values can range between (−0.999, 0.999), where a higher separation value indicates cluster structure with more separable clusters. Size of the dataset, number of clusters, and separation value of the clusters in the dataset is printed in file name. size_X_n_Y_sepval_Z.csv: Size of the dataset = X number of clusters in the dataset = Y separation value of the clusters in the dataset =